CN105293849A - Extraction method for tightly-bound extracellular polymeric substance (TB-EPS) in biological denitrification sludge - Google Patents
Extraction method for tightly-bound extracellular polymeric substance (TB-EPS) in biological denitrification sludge Download PDFInfo
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Abstract
The invention provides an extraction method for TB-EPS in biological denitrification sludge, which belongs to the field of biological treatment of sewage. The EPS in the biological denitrification sludge can be rapidly and highly-efficiently extracted by using an ultrasonic method under the conditions of ultrasonic intensity of 180 to 220 W and ultrasonic time of 5 to 20 min; and the optimal extraction method for proteins and polysaccharides in the TB-EPS in the biological denitrification sludge is provided, so technical parameters are provided for subsequent accurate quantification and research on the properties of the biological denitrification sludge in sewage treatment.
Description
Technical field
The present invention relates to field of biological sewage treatment, there is provided a kind of to combine closely type extracellular polymeric (TB-EPS) extracting method for biological denitrificaion mud, biological denitrificaion class mud extracellular polymeric can be extracted quickly and efficiently, accurate quantitative analysis for subsequent protein, polysaccharide and DNA provides the extracting method of stability and high efficiency, contributes to the research of biological denitrificaion sludge character in follow-up sewage treatment process.
Background technology
Extracellular polymeric (ExtracellularPolymericSubstances, EPS) is by microorganism under certain environment condition, mainly bacterium, secretes in some external high molecular polymers.Main component is similar to composition in the born of the same parents of microorganism, is some polymer substances, as polymkeric substance such as polysaccharide, protein and nucleic acid.Research report thinks that EPS is as the important component in active sludge except cell and moisture, it is mainly derived from the related component in the secretory product of microorganism cells metabolism, polymkeric substance that aqtocytolysis produces, the surface mass of cell detachment and water inlet matrix, and its content accounts for active sludge total organic matter 50% ~ 90%.
In recent years, having scholar to propose EPS divides layer component to divide: rete malpighii (slime), loose attachment EPS (looselyboundextracellularpolymericsubstances, and close adhesion EPS (tightlyboundextracellularpolymericsubstances, TB-EPS) LB-EPS).The component of EPS and content have important impact to the flocculation of active sludge, the processing property such as sedimentation and dehydration.EPS extracts institute's choosing method as far as possible exactly by it from cell surface extraction and isolation out, and will need gentle extracting method, avoids the interior polymers of cell to leak into outside born of the same parents, causes cracking and the dissolving of cell.To the standard of extraction usually using the relative content of EPS amplifying nucleic acid as inspection cytoclasis degree of Extracellular polymers (EPS).At present, the large class methods of physics and chemistry two are roughly divided into the extraction of EPS, but have different relative merits separately, such as: in numerous extracting method, formaldehyde-NaOH method is because having stronger extractability, but can have an impact to subsequent measurements, although after extraction EPS, dialysis method can be adopted to remove residual formaldehyde, but, dialysis procedure usually needs for a long time and easily causes EPS to lose, and increases error at measurment, so far to the extraction of mud EPS still without unified standard method.
In order to study the EPS of each layer of mud and component thereof better to the impact of Sludge Property, need to take to have more for different mud to optimize extracting method targetedly, exactly type of the combining closely extracellular polymeric (TB-EPS) in mud is extracted, thus each component concentration in accurate quantitative analysis EPS, evaluate sludge characteristics.
Summary of the invention
For the weak point of above-mentioned research, the invention provides and a kind of to combine closely type extracellular polymeric (TB-EPS) extracting method for biological denitrificaion class mud, biological denitrificaion class mud extracellular polymeric can be extracted quickly and efficiently, accurate quantitative analysis for subsequent protein, polysaccharide and DNA provides the extracting method of stability and high efficiency, so that the follow-up research carrying out biological denitrificaion sludge characteristics.
A kind of to combine closely type extracellular polymeric (TB-EPS) extracting method for biological denitrificaion class mud, it is characterized in that: comprise the following steps
In biological denitrificaion sludge system, mud mixed liquid suspended sediment concentration (MLSS) mean value is 2000-3000mg/L (preferred 2700mg/L), sludge volume index (SVI) mean value is 70-130mL/g (preferred 98mL/g), and system ammonia nitrogen removal frank is 100%; Ultrasound intensity is adopted to be 180-220W for combine closely type extracellular polymeric (TB-EPS) protein and Polyose extraction of this biological denitrificaion mud, the ultrasonic 20min of ultrasonic time 5min-20min, preferred 192W.
Above-mentioned biological denitrificaion mud is extracted the mud substrate 1*PBS solution after loose type extracellular polymeric (LB-EPS) and recovered original volume, then carried out ultrasonic.
Compared with prior art, the present invention has the following advantages:
(1) optimization method extracted for biological denitrificaion mud extracellular polymeric (EPS) provided by the invention is mainly for the extraction optimization method of type extracellular polymeric of combining closely in biological denitrificaion function mud, determines the optimum ultrasound intensity extracting type polymkeric substance of combining closely outside this type of mud born of the same parents.
(2) method mainly for extracting in biological denitrificaion function mud type extracellular polymeric of combining closely provided by the invention, under determining optimum ultrasound intensity condition, not serious broken born of the same parents extract the optimum ultrasonic time of type extracellular polymeric of combining closely, and provide guide parameters to the extraction of extracellular polymeric in this type of mud.
Accompanying drawing explanation
TB-EPS extraction effect figure under the different ultrasound intensity condition of Fig. 1;
TB-EPS extraction effect figure under the different ultrasonic time condition of Fig. 2.
Embodiment
Below in conjunction with accompanying drawing and specific implementation method, the present invention is described in further detail, but the present invention is not limited to following examples.
Embodiment 1
1, the basic parameter of biological denitrificaion class mud is determined: average mixed genetic-neural network (MLSS) is 2700mg/L, mean sludge bulk index (SVI) value is 98mL/g, system ammonia nitrogen removal frank is 100%, and total nitrogen average removal rate is 56.8% (system water inlet total nitrogen mean value is 71.6mg/L);
2, get 180ml test mud sample and remove the materials such as coarse sand by the mesh screen of 1.2mm, be divided into 12 groups, every part of 15ml mud sample;
3, the former mud sieved precipitates 1.5 hours at 4 DEG C, with siphon pipe, upper solution is collected as supernatant layer EPS;
4, by under throw out whizzer 2000g condition centrifugal 15 minutes, upper liquid is collected as slime layer EPS;
5, substrate 1*PBS solution recovers original volume, suspension under 5000g condition centrifugal 15 minutes, and upper liquid is collected as LB-EPS;
6, the mud substrate 1*PBS solution after loose type extracellular polymeric (LB-EPS) will have been extracted and recovered original volume;
7, adopt total power to be that the ultrasound intensity of 20%, 30%, 40%, 50%, 60% of 480w carries out 5min supersound process to 1-5 sample respectively, adopting the centrifugal 20min of 20000g rotating speed to collect upper liquid is the TB-EPS that under ultrasonic 5min condition, different ultrasound intensity extracts.
8, the ultrasound intensity of restraint test is 40%, adopt the ultrasonic time of 5min, 10min, 15min, 20min, 25min, 30min, 40min to carry out supersound process to 6-12 sample respectively, adopting the centrifugal 20min of 20000g rotating speed to collect upper liquid is the TB-EPS that under ultrasound intensity 40% condition, different ultrasonic time extracts.
9, by the membrane filtration of each group of TB-EPS extracted with 0.45 μm;
10, folin-phenol method protein graticule, anthrone method polysaccharide graticule and pentanoic method DNA graticule is set up, and protein, DNA and polysaccharide content in working sample;
11, the EPS obtained under different ultrasound intensity condition is extracted data results and carry out com-parison and analysis (Fig. 1), it is 40% that test obtains this biological denitrificaion mud optimum ultrasound intensity of type extracellular polymeric (TB-EPS) protein and Polyose extraction of combining closely, protein, polysaccharide and DNA content that the ultrasonic 5min time extracts are respectively 19.74,8.86 and 1.55mg/L, DNA percentage composition is 5.14%, does not occur seriously breaking born of the same parents;
12, the EPS obtained under different ultrasonic time condition is extracted data results and carry out com-parison and analysis (Fig. 2), it is that optimum ultrasonic time is 20min under 192W condition that test obtains for combine closely type extracellular polymeric (TB-EPS) protein and Polyose extraction ultrasound intensity of this biological denitrificaion mud, the protein extracted, polysaccharide and DNA content are respectively 148.05,14.91 and 10.34mg/L, DNA percentage composition is 5.97%, does not occur seriously breaking born of the same parents.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (5)
1. to combine closely type extracellular polymeric (TB-EPS) extracting method for biological denitrificaion class mud, it is characterized in that: comprise the following steps
In biological denitrificaion sludge system, mud mixed liquid suspended sediment concentration (MLSS) mean value is 2000-3000mg/L, and sludge volume index (SVI) mean value is 70-130mL/g, and system ammonia nitrogen removal frank is 100%; Adopt ultrasonic method for combine closely type extracellular polymeric (TB-EPS) protein and Polyose extraction of this biological denitrificaion mud, ultrasound intensity is 180-220W, ultrasonic time 5min-20min.
2. according to the method for claim 1, it is characterized in that, biological denitrificaion mud has been extracted the mud substrate 1*PBS solution after loose type extracellular polymeric (LB-EPS) and recovered original volume, then carried out ultrasonic.
3. according to the method for claim 1, it is characterized in that, in biological denitrificaion sludge system, mud mixed liquid suspended sediment concentration (MLSS) mean value is 2700mg/L, and sludge volume index (SVI) mean value is 98mL/g.
4. according to the method for claim 1, it is characterized in that, system water inlet total nitrogen mean value is 71.6mg/L.
5. according to the method for claim 1, it is characterized in that, ultrasound intensity is 192W, ultrasonic time 20min.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106525738A (en) * | 2016-10-24 | 2017-03-22 | 北京工业大学 | Method for measuring biomass in embedding gel |
| CN107417057A (en) * | 2017-05-08 | 2017-12-01 | 同济大学苏州研究院 | A kind of extracellular polymeric substances from activated sludge layering and extracting method |
| CN108569831A (en) * | 2018-04-25 | 2018-09-25 | 中原工学院 | A kind of extraction separation method of effective activated sludge exoprotein and application |
| CN109320627A (en) * | 2018-10-08 | 2019-02-12 | 西藏大学 | A kind of extracting method of biological sludge extracellular polymeric |
| CN114573095A (en) * | 2022-03-10 | 2022-06-03 | 西安建筑科技大学 | Biological denitrification, sludge reduction and hot water production increase treatment operation method and sludge thereof |
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| WO2002031184A2 (en) * | 2000-10-11 | 2002-04-18 | Universite De Liege | Method for extracting intact micro-organisms from sludge or biofilms |
| CN102781226A (en) * | 2009-12-07 | 2012-11-14 | 朱伟星 | Standardization of tissue specimen preservation by ultrasound and temperature control |
| CN104316502A (en) * | 2014-10-18 | 2015-01-28 | 北京工业大学 | Method for optimizing ultrasonic conditions and extracting extracellular polymeric substances through three-dimensional fluorescence spectrum |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2002031184A2 (en) * | 2000-10-11 | 2002-04-18 | Universite De Liege | Method for extracting intact micro-organisms from sludge or biofilms |
| CN102781226A (en) * | 2009-12-07 | 2012-11-14 | 朱伟星 | Standardization of tissue specimen preservation by ultrasound and temperature control |
| CN104316502A (en) * | 2014-10-18 | 2015-01-28 | 北京工业大学 | Method for optimizing ultrasonic conditions and extracting extracellular polymeric substances through three-dimensional fluorescence spectrum |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106525738A (en) * | 2016-10-24 | 2017-03-22 | 北京工业大学 | Method for measuring biomass in embedding gel |
| CN107417057A (en) * | 2017-05-08 | 2017-12-01 | 同济大学苏州研究院 | A kind of extracellular polymeric substances from activated sludge layering and extracting method |
| CN108569831A (en) * | 2018-04-25 | 2018-09-25 | 中原工学院 | A kind of extraction separation method of effective activated sludge exoprotein and application |
| CN109320627A (en) * | 2018-10-08 | 2019-02-12 | 西藏大学 | A kind of extracting method of biological sludge extracellular polymeric |
| CN114573095A (en) * | 2022-03-10 | 2022-06-03 | 西安建筑科技大学 | Biological denitrification, sludge reduction and hot water production increase treatment operation method and sludge thereof |
| CN114573095B (en) * | 2022-03-10 | 2023-09-26 | 西安建筑科技大学 | Biological denitrification, sludge reduction and increased heat production water treatment operation method |
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Application publication date: 20160203 |